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101.
Kristel Eggermont Inge J. Goderis Willem F. Broekaert 《Plant Molecular Biology Reporter》1996,14(3):273-279
We describe here a reliable high-throughput method for extraction of RNA from fresh or frozen plant tissue that obviates laborious
and time-consuming homogenisation by mortar and pestle. The method is based on homogenisation by high-speed reciprocal shaking
in presence of a mixture of inexpensive abrasive materials; i.e., quartz sand and glass beads. After homogenisation, the method
follows a standard procedure for RNA extraction by phenol/LiCl. Yield and quality of RNA obtained by homogenisation with the
sand/glass bead mix are identical to those obtained by mortar and pestle. 相似文献
102.
肾综合征出血热人体组织中病毒包涵体的性质及意义的进一步研究 总被引:2,自引:0,他引:2
应用免疫组化、原位分子杂交、电镜及免疫电镜等方法进一步对肾综合征出血热人体尸检组织中病毒包涵体(IB)的抗原、核酸性质和超微结构特点作了进一步观察。结果,在39例中的20例尸检病例组织中显示出病毒核蛋白抗原和血凝素抗抗原阳性的IB,其中包括16例陕西尸检病例组织中的6例休克期和1例多尿期病例,17例上海病例中的3例休克期和9例少尿期病例及3例江西病例中的1例休克期病例。IB主要分布在呼吸道和肺泡、肾远曲小管和集合管、胃肠道、腺垂体、扁桃体、胰腺、前列腺等组织的粘膜上皮和腺上皮细胞及肝细胞和睾丸生精上皮细胞胞浆中,阳性细胞形态基本正常。应用原位分子杂交,可在该组细胞小同时检测到病毒RNA,多为胞浆内弥漫阳性,仅少数组织中显示出病毒RNA阳性IB结构。电镜观察阳性组织细胞中出现由大量微丝微管及颗粒样结构组成的IB结构,其小的病毒颗粒状结构、内质网及纤维丝状结构呈病毒抗原阳性,上述结构位于高尔基体区。结果说明该病毒有感染上皮细胞的特性,对其宿主细胞的致细胞病变作用是极其温和的,且多表现在亚细胞水平。IB可能是病毒过量表达抗原的堆积或病毒复制部位,而微丝微管结构可能参与病毒的感染过程。 相似文献
103.
Manfred Tacker Walter Fontana Peter F. Stadler Peter Schuster 《European biophysics journal : EBJ》1994,23(1):29-38
We present and study the behavior of a simple kinetic model for the melting of RNA secondary structures, given that those structures are known. The model is then used as a map that. assigns structure dependent overall rate constants of melting (or refolding) to a sequence. This induces a landscape of reaction rates, or activation energies, over the space of sequences with fixed length. We study the distribution and the correlation structure of these activation energies.
Correspondence to: P. Schuster 相似文献
104.
105.
Desmond R. Jimenez Jeffrey P. Shapiro Raymond K. Yokomi 《Entomologia Experimentalis et Applicata》1994,70(2):143-152
This study was conducted to evaluate the effect of two different biotypes of the sweetpotato whitefly,Bemisia tabaci (Gennadius), on the induction of squash silverleaf (SSL), and to determine if double-stranded RNA (dsRNA) occurs in geographically
remote populations of the two biotypes. Recently collected B-biotype whiteflies from Florida, Arizona, Mississippi, and Texas
(SPW-B) all contained a 7.0 kb dsRNA molecule. Kb dsRNA molecule. Laboratory colonies of A-biotype whiteflies that were originally
collected in 1981 from cotton in Arizona and California did not contain the 7.0 Kb dsRNA. When the two biotypes were compared
only the SPW-B induced rapid onset, grade 5, SSL. DsRNA similar to that found in adult SPW-B was concentrated in whitefly
nymphs, but host plant leaf tissue did not contain any consistent dsRNA molecules. SPW-A only induced low-grade SSL and progeny
of SPW-A that were fed on pumpkin plants displaying SSL did not acquire the ability to express dsRNA or induce SSL. Our data
suggest that dsRNA is not directly involved in the induction of SSL and that SSL is a host-specific response, to a feeding
injury induced by B-biotype whiteflies. The origin and source of the 7.0 Kb dsRNA molecule remains enigmatic but its expression
is constant in the whitefly biotype that is responsible for the induction of SSL and several other plant disorders in the
U.S. 相似文献
106.
Evolution of secondary structure in the family of 7SL-like RNAs 总被引:8,自引:0,他引:8
Primate and rodent genomes are populated with hundreds of thousands copies of Alu and B1 elements dispersed by retroposition, i.e., by genomic reintegration of their reverse transcribed RNAs. These, as well as primate BC200 and rodent 4.5S RNAs, are ancestrally related to the terminal portions of 7SL RNA sequence. The secondary structure of 7SL RNA (an integral component of the signal recognition particle) is conserved from prokaryotes to distant eukaryotic species. Yet only in primates and rodents did this molecule give rise to retroposing Alu and B1 RNAs and to apparently functional BC200 and 4.5S RNAs. To understand this transition and the underlying molecular events, we examined, by comparative analysis, the evolution of RNA structure in this family of molecules derived from 7SL RNA.RNA sequences of different simian (mostly human) and prosimian Alu subfamilies as well as rodent B1 repeats were derived from their genomic consensus sequences taken from the literature and our unpublished results (prosimian and New World Monkey). RNA secondary structures were determined by enzymatic studies (new data on 4.5S RNA are presented) and/or energy minimization analyses followed by phylogenetic comparison. Although, with the exception of 4.5S RNA, all 7SL-derived RNA species maintain the cruciform structure of their progenitor, the details of 7SL RNA folding domains are modified to a different extent in various RNA groups. Novel motifs found in retropositionally active RNAs are conserved among Alu and B1 subfamilies in different genomes. In RNAs that do not proliferate by retroposition these motifs are modified further. This indicates structural adaptation of 7SL-like RNA molecules to novel functions, presumably mediated by specific interactions with proteins; these functions were either useful for the host or served the selfish propagation of RNA templates within the host genome.Abbreviations FAM
fossil Alu element
- FLAM
free left Alu monomer
- FRAM
free right Alu monomer
- L-Alu
left Alu subunit
- R-Alu
right Alu subunit
Correspondence to: D. LabudaDedicated to Dr. Robert Cedergren on the occasion of his 25th anniversary at the University of Montreal 相似文献
107.
Selenite has been shown to undergo intracellular metabolism that results in its conversion to other low molecular weight Secontaining
species and also to its incorporation into a selenocysteine residue in selenoprotein. In order to investigate whether the
incorporation into protein is required for the cytotoxic effects of selenite, we have examined whether inhibition of protein
synthesis prevents the inhibitory effect of selenite on the ability of cells to form colonies or to synthesize RNA. We have
found that treatment of HeLa cells with cycloheximide inhibited protein synthesis by >90% but had no effect on the inhibitory
effect of selenite on cell colony formation or RNA synthesis. Since protein synthesis is not necessary for these cytotoxic
effects of selenite they are unlikely to result from an increase in the synthesis of selenoproteins. 相似文献
108.
109.
110.
Abstract: Derivatives of the lac promoter (tac, pac, rac) belong to the strongest bacterial promoters which are frequently used for the induced overexpression of foreign genes in Escherichia coli . However, their use in fermentation processes is strongly restricted because of the high cost of the inducer iso-propyl-β-D-thiogalactopyranoside (IPTG). The aim of this work was to investigate the possibility of using lac-derived promoters in high cell density processes resulting in a high yield of the induced recombinant protein if glucose is the main carbon and energy source. Lactose is tested as inducer of the main antigenic coat protein (VP1) of the foot and mouth disease (FMD) virus in a T7-RNA polymerase expression system. It was shown that lactose is able to induce the expression of the recombinant gene to an amount of the VP1 protein corresponding to 20% of the total cell protein. 相似文献